Hi netters,
I have been having problems for extracting genomic DNA from cultures of
lactic acid bacteria (Lactobacillus fermentum). I have tried the
guanidium thiocyanate method, but the outcome has been degraded DNA in
low yields. A lot of "dna" is formed when ethanol precipitating, but most
of it remains undissolved in the TE solution at the end. We have tried a
lot of variations, like overnight lisozyme, previous proteinase K
digestion, more or less chloroform extractions, more or less time and
temperature of
incubation with the guanidium solution, checked pH of all reagents, etc.
Has anybody got a tailor-made protocol for extracting genomic DNA for PCR
and other molecular works with Lactobacillus fermentum (or relatives)?
Thanks for any help.
Cheers from the tropics,
Heitor L.C. Coutinho
Funda at ao Tropical Andre Tosello
Campinas - SP, Brazil